Multimodality labeling strategies for the investigation of nanocrystalline cellulose biodistribution in a mouse model of breast cancer

Methods We have developed a nuclear and fluorescence labeling strategy for nanocrystalline cellulose (CNC), an emerging biomaterial with versatile chemistry and facile preparation from renewable sources. We modified CNC through 1,1′-carbonyldiimidazole (CDI) activation with radiometal chelators desferrioxamine B and 1,4,7-triazacyclononane-1,4,7-triacetic acid (NOTA), allowing for the labeling with zirconium-89 (t½ = 78.41 h) and copper-64 (t½ = 12.70 h), respectively, for non-invasive positron emission tomography (PET) imaging. The far-red fluorescent dye Cy5 was added for ex vivo optical imaging, microscopy and flow cytometry. The multimodal CNC were evaluated in the syngeneic orthotopic 4T1 tumor model of human stage IV breast cancer. Results Modified CNC exhibited low cytotoxicity in RAW 264.7 macrophages over 96 h, and high radiolabel stability in vitro. After systemic administration, radiolabeled CNC were rapidly sequestered to the organs of the reticulo-endothelial system (RES), indicating immune recognition and no passive tumor targeting by the enhanced permeability and retention (EPR) effect. Modification with NOTA was a more favorable strategy in terms of radiolabeling yield, specific radioactivity, and both the radiolabel and dispersion stability in physiological conditions. Flow cytometry analysis of Cy5-positive immune cells from the spleen and tumor corroborated the uptake of CNC to phagocytic cells. Conclusions Future studies on the in vivo behavior of CNC should be concentrated on improving the nanomaterial stability and circulation half-life under physiological conditions and optimizing further the labeling yields for the multimodality imaging strategy presented. Advances in knowledge Our studies constitute one of the first accounts of a multimodality nuclear and fluorescent probe for the evaluation of CNC biodistribution in vivo and outline the pitfalls in radiometal labeling strategies for future evaluation of targeted CNC-based drug delivery systems. Implications for patient care Quantitative and sensitive molecular imaging methods provide information on the structure–activity relationships of the nanomaterial and guide the translation from in vitro models to clinically relevant animal models.Peer reviewe

Mir-135b induces osteosarcoma invasion by the modulation of foxo-1 and c-myc

Background: Osteosarcoma (OS) is the most common type of bone malignancy. Many studies have attempted to find the association between microRNAs and cancer-associated processes. Alterations in miRNA expression through genetic or epigenetic changes, impairment of transcription factors, and ectopic expression of miRNAs induce the development and progression of cancer. Al-though miR-135b has been thoroughly documented as an oncogene in the majority of studies, some controversies remain about the conflicting role of miR-135b as a tumor-suppressor. Objectives: The present study aimed at investigating the oncogenic and/or tumor-suppressing role of miR-135b in human OS. Methods: In this study, 21 OS tissue samples, along with 21 adjacent bone tissues (normal) as control specimens were collected to analyze the expression of miR-135b. The Saos2 cell-line was transiently transfected with the miR-135b mimic and inhibitor to assess its effect on two critical transcription factors, namely FOXO-1 and c-Myc. qRT-PCR was performed to quantify the expression of miR-135b in both OS tissues and the Saos2 cell-line. The MTT, cell migration, and cell invasion assays were used to characterize the miR-135b function. The western blot analysis was carried out to monitor the targets of miR-135b. Finally, the changes in cellular functions such as migration and invasion, following the transfection of miR-135b mimic and inhibitor, were verified. Results: The results showed that in comparison with the adjacent normal bone tissues, the expression of miR-135b was higher in OS tissue samples, which inversely correlated with the expression rate of FOXO-1, whereas the expression of c-Myc had a direct relationship to miR-135b expression. Functionally, the miR-135b mimic led to an increase in cell proliferation, invasion, and migration of OS cancer cells. Conclusions: MiR-135b induces the proliferation and invasion of OS cells by the degradation of FOXO-1 and upregulation of c-Myc. © 2020, Author(s)

Exogenous coenzyme Q10 modulates MMP-2 activity in MCF-7 cell line as a breast cancer cellular model

<p>Abstract</p> <p>Background/Aims</p> <p>Matrix Metalloproteinases 2 is a key molecule in cellular invasion and metastasis. Mitochondrial ROS has been established as a mediator of MMP activity. Coenzyme Q₁₀contributes to intracellular ROS regulation. Coenzyme Q₁₀beneficial effects on cancer are still in controversy but there are indications of Coenzyme Q₁₀complementing effect on tamoxifen receiving breast cancer patients.</p> <p>Methods</p> <p>In this study we aimed to investigate the correlation of the effects of co-incubation of coenzyme Q10 and N-acetyl-L-cysteine (NAC) on intracellular H2O2 content and Matrix Metalloproteinase 2 (MMP-2) activity in MCF-7 cell line.</p> <p>Results and Discussion</p> <p>Our experiment was designed to assess the effect in a time and dose related manner. Gelatin zymography and Flowcytometric measurement of H2O2 by 2'7',-dichlorofluorescin-diacetate probe were employed. The results showed that both coenzyme Q10 and N-acetyl-L-cysteine reduce MMP-2 activity along with the pro-oxidant capacity of the MCF-7 cell in a dose proportionate manner.</p> <p>Conclusions</p> <p>Collectively, the present study highlights the significance of Coenzyme Q₁₀effect on the cell invasion/metastasis effecter molecules.</p

Production of Mouse Monoclonal Antibody against Morphine without Cross Reactivity with Heroin

Introduction: Immunoassay procedures for detecting and determining opioids in blood and other biologic fluids are based on Monoclonal antibodies. In the present study, monoclonal antibody against Morphine was taken into account. Methods: Hybridoma protocol was used in order to produce the monoclonal antibody against morphine in mice. For this purpose, five 6–8-week old female BALB/c mice were immunized with morphine C6- hemisuccinated derivative conjugated to cationized bovine serum albumin (cBSA). The spleens lymphocytes were fused with SP2/0 cells using polyethylene glycol (PEG). Hybridoma clones were subcloned by limiting dilution. Class and subclass of monoclonal antibody were determined using Roche isostrip test. Moreover, antibody was purified by protein G affinity chromatography and affinity was determined according to the method described by Beatty et al. Finally, the cross reaction of monoclonal antibody was determined with some structurally related molecules such as codeine and apomorphine. Results: Among 3 hybridoma clones that reacted with the morphine-BSA, but not with BSA, after thrice limiting dilution, one stable hybridoma monoclon was obtained. The monoclonal antibody (MAb) was found to be of IgG2b class and subclass and containing lambda light chain. The affinity of the MAb to morphine was obtained 2.8 ×109 M-1 by non competitive enzyme immunoassay. The titer of supernatant of cell culture medium was 1/400. The MAb was cross reacted with codeine (100%) and apomorphine (16.5%), though no reaction was observed with heroin, naloxone, naltrexone, and papavrin. Conclusion: The study findings revealed that the produced antibody against morphine was comparable with other antibodies for specificity and affinity; therefore it is usable in design of diagnostic immunoassay in biologic fluids

Evaluation of periodontal condition of menopause women with osteoporosis and osteopenia and comparison with control group

Background and Aim: Several risk factors directly affect the development of periodontal diseases. Also some systemic diseases act indirectly as predisposing and aggrevating factors. Osteoporosis is one of these factors and one of its main causes is lack of physical activity in postmenopause period. The incidence of osteoporosis is increasing in our country. The goal of this study was to evaluate the periodontal condition of women with osteoporosis and osteopenia referred to bone densitometric division of Loghman hospital in 2003 and compare to control group. Materials and Methods: In this case control study based on BMD (Bone Mineral Density) measurement of back and thigh using DEXA method, 60 patients referred to bone densitometric division of Loghman hospital, were randomly selected. Cases were divided into three groups, 20 with osteoporosis, 20 with osteopenia and 20 normal cases. Periodontal indices consisting of plaque index (PI), tooth loss (TL), gingival recession (GR), probing pocket depth (PPD) and papilla bleeding index (PBI) were evaluated by clinical and radiographic examination. Data were analyzed by Kruskall Wallis and Dunn tests with p&lt;0.05 as the limit of significance. Results: PBI, PI and TL were significantly higher in osteoporotic group than osteopenic and normal group. PPD was not different in the three groups. Due to the low prevalence of recession in our study, this parameter was not included in the statistical analysis. Conclusion: It seems that osteoporosis does not increase the incidence of periodontal diseases because it affects bone quality rather than quantity. In osteoporosis calcium deficiency and increasing age lead to decreased physical activity and ultimately affect the patient&apos;s oral hygiene performance. Thus, periodontal manifestations are presented as gingival bleeding and gingivitis

Histologic and histomorphometric study on the effects of lactoferrin and porous bovine bone mineral (Bio-Oss) on the regeneration of bone defects made on rabbit calvarium

&quot;nBackground and Aims: Nowadays reconstruction of alveolar defects has become one of dentists&apos; problems especially in areas which are going to get dental implants. Inorganic bovine bone mineral (Bio-Oss) is one of the most popular graft materials that acts as a structure for migration of osteoblasts. If migration, proliferation, and differentiation of osteoblasts can be promoted by a material, it would be possible to reconstruct more amount of bone in a shorter period of time. Milk contains vital proteins that regulate bone growth. One of these important proteins is lactoferrin. The aim of this study was to examine the effect of added bovine lactoferrin to Bio-Oss on osteogenesis.&quot;nMaterials and Methods: Two doses of 50 and 500 &amp;micro;g/ml of lactoferrin were prepared. Ten New Zealand white rabbits were selected for this study. Four 6-mm symmetrical detects were created in each rabbit&apos;s calvarium. Two of these sites were filled with Bio-Oss that was wetted with two doses of lactoferrin. Third detect was filled with Bio-Oss alone and the forth one was left empty as control group. After 4 weeks histologic and histomorphometric analysis was performed.&quot;nResult: There was no sign of obvious inflammation in any of four groups. Also there was no difference among four groups in terms of vitality, type of new bone, and foreign body reaction. However, amount of bone formation in control group was significantly lower compared with the other 3 groups. Although lactoferrin containing groups showed little increase in bone formation especially in higher concentration, there was not statistically significant difference among the three test groups. Amount of remaining biomaterial also was lower in lactoferrin containing groups compared with the Bio-Oss group but the differences were not significant.&quot;nConclusion: Although there was no significant difference among the test groups, it seems that the added lactoferrin increases bone formation. Considering the limitations of this study, more studies are needed in different concentrations of lactoferrin and different healing periods. Furthermore, because of possible washout of the lactoferrin from the defects, it would be helpful to find and evaluate a proper carrier agent for lactoferrin to see its real effects

A comparative histologic and histomorphometric investigation on Biostite in surgical defects

Statement of Problem: The goal of bone regenerative therapy is to reconstruct the damaged tissues to initial condition, which seems so important in periodontoiogy and implantology. In such therapies, different biomaterials, with some advantages and disadvantages, may by use.&quot;nPurpose: The aim of this research was to evaluate the quality, bone trabecular density and thickness of newly forming bone as well as foreign body reaction against Biostite&amp;reg;, in comparison with the control group.&quot;nMaterial and Methods: In this experimental study the total number of 5 sheep was operated with&quot;ngeneral anesthesia. At first 10 standard defects were created in 5x5x5mm on the left and right edentulous&quot;nmandibular ridge. Five defects were randomly treated by Biostite&amp;reg;, the remaining five were considered&quot;nas controls. After six months, the sheep were sacrificed and the separated defect areas with intact margin&quot;nwere placed in 10% formaline and prepared for histologic and histomorphometric study. To analyze the&quot;ndata, Mann-Whitney U test was used.&quot;nResults: The mean percentage of new bone formation in Biostite&amp;reg; group was statistically more than the&quot;ncontrol group (PO.05). Newly formed bone was vital in both groups which was a combination of&quot;nlamellar and woven types, however, the thickness and density of new bone in biostite&amp;reg; group was more&quot;nthan the controls. Foreign body reaction was observed in Biostite&amp;reg; group.&quot;nConclusion: It is suggested that Biostite&amp;reg; synthetic material can be useful in bone regenerative&quot;ntherapies

Surveying the Relation of Shahed Stu-dents’ Cognition of Academic Atmos-phere, and Their Individual and Family characteristics with Social Compatibility

Determining the relation of Shahed students’ cognition of academic at-mosphere and their individual and family characteristics with social compatibility is purpose of this study. Subjects are 315 male and female Shahed students, selected among all of Tehran universities’ students us-ing stratified sampling with appropriate allocation. The applied tools in this research are California personality questionnaire and structured questionnaire for measurement of Shahed students’ cognition of academ-ic atmosphere and Shahed students’ demographic features. The data were analyzed by hierarchical regression. Findings showed that Stu-dents’ satisfaction from Shahed privilege, other students’ positive view-point to Shahed students, professors’ attitude, and marriage of Shahed students are predictors of Shahed students’ social compatibility. No meaningful relation was observed between students’ family variables such as their mothers’ remarriage and social compatibility. Possible rea-sons for lack of meaningful relation between family variable variables such as their mothers’ remarriage and social compatibility was discussed